Artesunate enhances the antibacterial effect of [beta]-lactam antibiotics against Escherichia coli by increasing antibiotic accumulation via inhibition of the multidrug efflux pump system AcrAB-TolC.
Li, Bin 1,+; Yao, Qi 1,+; Pan, Xi-Chun 1; Wang, Ning 2; Zhang, Rong 1; Li, Jun 1; Ding, Guofu 1; Liu, Xin 2; Wu, Chong 1; Ran, Dongzhi 1; Zheng, Jiang 2; Zhou, Hong 1,*
[Article]
Journal of Antimicrobial Chemotherapy.
66(4):769-777, April 2011.
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Objectives: Occasionally, we found that artesunate enhanced the antibacterial effects of antibiotics in vitro. Therefore, the enhancement of various [beta]-lactam antibiotics by artesunate against Escherichia coli and the possible mechanism were investigated in the present study.
Methods: Antibacterial effects were observed using the serial 2-fold dilution method and dynamic bacterial growth. Daunomycin accumulation within E. coli was observed using fluorospectrophotometry and laser confocal scanning microscopy. AcrAB-TolC, AmpC and TEM-1 mRNA expression was observed using a PCR method. Antisense oligonucleotides (as-ODNs) targeting AcrB were designed and used to block AcrB gene expression within E. coli ATCC 35218.
Results: Although artesunate itself had no antibacterial ability, artesunate significantly increased the antibacterial effect of [beta]-lactam antibiotics against E. coli ATCC 35218 and an E. coli clinical strain. Artesunate increased daunomycin accumulation within E. coli ATCC 35218 in a dose-dependent manner and reduced the mRNA expression of AcrAB-TolC, an important multidrug efflux system for Gram-negative bacteria. The bacterial number was significantly reduced by as-ODN targeting AcrB, but did not further decrease after additional artesunate treatment. In contrast, artesunate lost its enhancement of [beta]-lactam antibiotics against E. coli AG100A, a strain lacking the gene encoding AcrAB, and artesunate did not increase daunomycin accumulation within E. coli AG100A. After the transformation of pET28a-AcrB into E. coli AG100A, artesunate regained enhancement of [beta]-lactam antibiotics. Furthermore, artesunate did not inhibit the expression of AmpC and TEM-1 mRNA.
Conclusions: Artesunate enhances the antibacterial effect of various [beta]-lactam antibiotics against E. coli, which might be associated with the suppression of a major multidrug resistance system, AcrAB-TolC.
(C) British Society for Antimicrobial Chemotherapy 2011. Published by Oxford University Press. All rights reserved.